Electroinduced Extraction of Human Ferritin Heavy Chain Expressed in Hansenula polymorpha

Ganeva, Valentina; Galutzov, B.; Angelova, Boyana; Suckow, Manfred

doi:10.1007/s12010-017-2627-9

Cited by 8 publications

(5 citation statements)

References 44 publications

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“…In some cases, yeast expression systems have been shown to work more efficiently for the biotechnical production of mammalian proteins with pharmaceutical relevance. Common hosts are S. cerevisiae , P. pastoris and Hansenula polymorpha [ 273 , 274 , 275 ]. However, filamentous fungi are still used for the heterologous production of different mammalian proteins, for instance, the human hormone peptide obestatin using recombinant T. reesei strains [ 276 ].…”

Section: Recombinant Dna Strategiesmentioning

confidence: 99%

Strategies for the Development of Industrial Fungal Producing Strains

Salazar-Cerezo

Vries

Garrigues

2023

JoF

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The use of microorganisms in industry has enabled the (over)production of various compounds (e.g., primary and secondary metabolites, proteins and enzymes) that are relevant for the production of antibiotics, food, beverages, cosmetics, chemicals and biofuels, among others. Industrial strains are commonly obtained by conventional (non-GMO) strain improvement strategies and random screening and selection. However, recombinant DNA technology has made it possible to improve microbial strains by adding, deleting or modifying specific genes. Techniques such as genetic engineering and genome editing are contributing to the development of industrial production strains. Nevertheless, there is still significant room for further strain improvement. In this review, we will focus on classical and recent methods, tools and technologies used for the development of fungal production strains with the potential to be applied at an industrial scale. Additionally, the use of functional genomics, transcriptomics, proteomics and metabolomics together with the implementation of genetic manipulation techniques and expression tools will be discussed.

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Section: Recombinant Dna Strategiesmentioning

confidence: 99%

Strategies for the Development of Industrial Fungal Producing Strains

Salazar-Cerezo

Vries

Garrigues

2023

JoF

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“…Recently, working with the thermotolerant yeast Hansenula polymorpha, we obtained maximal protein extraction at electrical conditions where outlet temperatures of 59-60 • C had been reached (Ganeva et al, 2018). It could be assumed that the effect of the temperature on the protein release efficiency depends on the system particularity-the cell wall composition and structure, the plasma membrane composition, the thermostability of the soluble proteins.…”

Section: Influence Of Cell Concentrationmentioning

confidence: 99%

“…The thiol compound, which increases cell wall porosity by reducing the disulfide bonds between the cell wall mannoproteins (Zlotnik et al, 1984), significantly enhanced the yield. As diffusion is a temperature-dependent process, we tried to accelerate the protein release by incubation of the cells at a higher temperature, as done earlier for the extraction of intracellular proteins (Ganeva et al, 2003(Ganeva et al, , 2018. The increase of the temperature from 24 to 30 • C had some impact on the rate and efficiency of protein release mainly during the first several hours, however, the maximal yield reached after 14-16 h of incubation at these two temperature was rather similar.…”

Section: Influence Of Cell Concentrationmentioning

confidence: 99%

“…Since the treated cells become more sensitive to lytic enzymes (Ganeva et al, 2014(Ganeva et al, , 2018 and to mechanical disintegration (Shynkaryk et al, 2009), the residual biomass after removal of the protein, could be further processed to induce autolysis (Monch and Stute, 2002;Maza et al, 2020) or it could be disintegrated by high pressure homogenization for recovery of additional valuable compounds-amino acids, nucleotides, proteins, and polysaccharides.…”

Section: Nucleic Acid Content Of the Extractsmentioning

confidence: 99%

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Extraction of Proteins and Other Intracellular Bioactive Compounds From Baker’s Yeasts by Pulsed Electric Field Treatment

Ganeva

Angelova

Galutzov

et al. 2020

Front. Bioeng. Biotechnol.

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Yeasts are rich source of proteins, antioxidants, vitamins, and other bioactive compounds. The main drawback in their utilization as valuable ingredients in functional foods and dietary supplements production is the thick, indigestible cell wall, as well as the high nucleic acid content. In this study, we evaluated the feasibility of pulsed electric field (PEF) treatment as an alternative method for extraction of proteins and other bioactive intracellular compounds from yeasts. Baker’s yeast water suspensions with different concentration (12.5–85 g dry cell weight per liter) were treated with monopolar rectangular pulses using a continuous flow system. The PEF energy required to achieve irreversible electropermeabilization was significantly reduced with the increase of the biomass concentration. Upon incubation of the permeabilized cells in water, only relatively small intracellular compounds were released. Release of 90% of the free amino acids and low molecular UV absorbing compounds, 80% of the glutathione, and ∼40% of the total phenol content was achieved about 2 h after pulsation and incubation of the suspensions at room temperature. At these conditions, the macromolecules (proteins and nucleic acids) were retained largely inside. Efficient protein release (∼90% from the total soluble protein) occurred only after dilution and incubation of the permeabilized cells in buffer with pH 8–9. Protein concentrates obtained by ultrafiltration (10 kDa cut off) had lower nucleic acid content (protein/nucleic acid ratio ∼100/4.5) in comparison with cell lysates obtained by mechanical disintegration. The obtained results allowed to conclude that PEF treatment can be used as an efficient alternative approach for production of yeast extracts with different composition, suitable for application in food, cosmetics and pharmaceutical industries.

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“…Meanwhile, yeast expression systems have been shown to be superior for the biotechnical production of mammalian proteins with pharmaceutical relevance. Most common host organisms are Saccharomyces cerevisiae , Pichia pastoris , and Hansenula polymorpha (Ganeva et al 2017; Mallu et al 2016; Shibui et al 2013). Nevertheless, filamentous fungi are still used for the heterologous production of different mammalian proteins, for example, the human hormone peptide obestatin was successfully produced using recombinant T. reesei strains (Sun et al 2016).…”

Section: Production Of Heterologous Productsmentioning

confidence: 99%

Inducible promoters and functional genomic approaches for the genetic engineering of filamentous fungi

Kluge

Terfehr

Kück

2018

Appl Microbiol Biotechnol

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In industry, filamentous fungi have a prominent position as producers of economically relevant primary or secondary metabolites. Particularly, the advent of genetic engineering of filamentous fungi has led to a growing number of molecular tools to adopt filamentous fungi for biotechnical applications. Here, we summarize recent developments in fungal biology, where fungal host systems were genetically manipulated for optimal industrial applications. Firstly, available inducible promoter systems depending on carbon sources are mentioned together with various adaptations of the Tet-Off and Tet-On systems for use in different industrial fungal host systems. Subsequently, we summarize representative examples, where diverse expression systems were used for the production of heterologous products, including proteins from mammalian systems. In addition, the progressing usage of genomics and functional genomics data for strain improvement strategies are addressed, for the identification of biosynthesis genes and their related metabolic pathways. Functional genomic data are further used to decipher genomic differences between wild-type and high-production strains, in order to optimize endogenous metabolic pathways that lead to the synthesis of pharmaceutically relevant end products. Lastly, we discuss how molecular data sets can be used to modify products for optimized applications.

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Electroinduced Extraction of Human Ferritin Heavy Chain Expressed in Hansenula polymorpha

Cited by 8 publications

References 44 publications

Strategies for the Development of Industrial Fungal Producing Strains

Strategies for the Development of Industrial Fungal Producing Strains

Extraction of Proteins and Other Intracellular Bioactive Compounds From Baker’s Yeasts by Pulsed Electric Field Treatment

Inducible promoters and functional genomic approaches for the genetic engineering of filamentous fungi

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